Using Amplified Fragment Length Polymorphism Analysis to Differentiate Isolates of Pasteurella Multocida Serotype 1

Aims: To examine the variability among Pasteurella multocida strains isolated from pigs (nasal, tonsil and lung specimens) and humans in France. Methods and Results: The genetic diversity of 117 French isolates of P. multocida, obtained from pigs (n = 101) and humans (n = 16) and three reference strains, was evaluated by pulsed‐field gel electrophoresis (PFGE) after macrorestriction with ApaI. Sixty‐four patterns were detected. The genetic relationships revealed five clusters (Aa1, Aa2, Aa3, Ab and B). The pig isolates obtained from pneumonic lungs and nasal cavities were clustered in groups Ab and Aa1, respectively (P < 0·05). Up to four different PFGE patterns were detected in the same farm. Isolates producing dermonecrotic toxins were clustered only in group Aa1, suggesting that the toxigenic isolates were more genetically homogenous than the others. Conversely, cluster Aa3 was significantly associated with human isolates even if the human isolates are spread over most of the clusters. Conclusions: Pasteurella multocida strains were genetically diverse, but pig and human isolates were significantly clustered in distinct phylogenetic groups. Significance and Impact of the Study: The discrimination index was >0·95 in both populations of human and pig isolates. Therefore, ApaI‐PFGE seems to be a useful tool for epidemiological tracing of P. multocida infections.

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“…2004), amplified fragment length polymorphism (Moreno et al. 2003; Blehert et al. 2008), multilocus sequence analysis (Davies et al.…”

Section: Introductionmentioning

confidence: 99%

Molecular diversity of porcine and human isolates ofPasteurella multocida

Marois

Fablet

Gaillot³

et al. 2009

Journal of Applied Microbiology

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“…However, a one-band difference resulted in a new profile number assignment. Considering P. multocida diversity and how quickly it has been shown to change within even a single outbreak, 3 REA profiles become less informative. In contrast, WGS was able to show relatedness of samples, which provided much more prospective epidemiologic data than REA.…”

Section: Discussionmentioning

confidence: 99%

Comparison of whole genome sequencing to restriction endonuclease analysis and gel diffusion precipitin-based serotyping of Pasteurella multocida

et al. 2017

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The gel diffusion precipitin test (GDPT) and restriction endonuclease analysis (REA) have commonly been used in the serotyping and genotyping of Pasteurella multocida. Whole genome sequencing (WGS) and single nucleotide polymorphism (SNP) analysis has become the gold standard for other organisms, offering higher resolution than previously available methods. We compared WGS to REA and GDPT on 163 isolates of P. multocida to determine if WGS produced more precise results. The isolates used represented the 16 reference serovars, isolates with REA profiles matching an attenuated fowl cholera vaccine strain, and isolates from 10 different animal species. Isolates originated from across the United States and from Chile. Identical REA profiles clustered together in the phylogenetic tree. REA profiles that differed by only a few bands had fewer SNP differences than REA profiles with more differences, as expected. The GDPT results were diverse but it was common to see a single serovar show up repeatedly within clusters. Several errors were found when examining the REA profiles. WGS was able to confirm these errors and compensate for the subjectivity in analysis of REA. Also, results of WGS and SNP analysis correlated more closely with the epidemiologic data than GDPT. In silico results were also compared to a lipopolysaccharide rapid multiplex PCR test. From the data produced in our study, WGS and SNP analysis was superior to REA and GDPT and highlighted some of the issues with the older tests.

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“…Candidate species include those with breeding ranges that extend into the eastern Canadian Arctic and with wintering ranges or migratory routes that include the Canadian east coast (e.g., Great Black-backed Gulls, Glaucous Gulls). However, without further genotypic analysis (e.g., Blehert et al 2008;Subaaharan et al 2010) of isolates collected from Atlantic Canada, the Canadian Arctic, the St. Lawrence Estuary, and other historical outbreaks across Canada to evaluate possible sources of the recent emergence of avian cholera in Canada's Atlantic coast as well as the eastern Arctic, we can only speculate about the origin of this outbreak.…”

Section: Discussionmentioning

confidence: 99%